Difference between revisions of "Part:BBa K525405:Design"

 
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__NOTOC__
 
__NOTOC__
 
<partinfo>BBa_K525405 short</partinfo>
 
<partinfo>BBa_K525405 short</partinfo>
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===Design Notes===
 
===Design Notes===
−
Fusion Protein of BBa_K525403 and BBa_J18931
 
  
 +
* <partinfo>K525403</partinfo> fused to <partinfo>J18931</partinfo>
 +
** <partinfo>K525403</partinfo> synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
 +
 +
* NgoMIV restriction site downstream the start codon to easily fuse an N-terminal domain to the coding sequence
 +
 +
* AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence
  
  
 
===Source===
 
===Source===
  
−
Fusion Protein of S-Layer sbpA and mCitrine  
+
* <partinfo>K525403</partinfo> fused to <partinfo>J18931</partinfo>
 +
** <partinfo>K525403</partinfo> synthesized, codon optimized and in Freiburg assembly standard (RFC 25) to easily create fusion proteins
 +
 
 +
* Promoter: fusion promoter between T7 promoter and lac-operator
 +
** T7 promoter from T7 phage
 +
** lac-operator from ''E. coli''
 +
 
 +
* <partinfo>K525401</partinfo> S-layer gene ''sbpA'' synthesized, originated in ''Lysinbacillus sphaericus''
 +
 
 +
* <partinfo>J18931</partinfo> yellow fluorescent protein (YFP) mCitrine
 +
** from parts.igem
 +
** was synthesized before it was sent in
 +
 
  
 
===References===
 
===References===
 +
Kainz B, Steiner K, Möller M, Pum D, Schäffer C, Sleytr UB, Toca-Herrera JL (2010) Absorption, Steady-State Fluorescence, Fluorescence Lifetime, and 2D Self-Assembly Properties of Engineered Fluorescent S-Layer Fusion Proteins of ''Geobacillus stearothermophilus'' NRS 2004/3a, [http://pubs.acs.org/doi/abs/10.1021/bm901071b ''Biomacromolecules'' 11(1):207-214].
 +
 +
Sleytr UB, Huber C, Ilk N, Pum D, Schuster B, Egelseer EM (2007) S-layers as a tool kit for nanobiotechnological applications, [http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2006.00573.x/full ''FEMS Microbiol Lett'' 267(2):131-144].

Revision as of 15:56, 13 September 2011

Fusion Protein of S-Layer SbpA and mCitrine


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 104
    Illegal BglII site found at 221
    Illegal XhoI site found at 1996
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 76
    Illegal AgeI site found at 3913
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 493
    Illegal BsaI.rc site found at 622


Design Notes

  • NgoMIV restriction site downstream the start codon to easily fuse an N-terminal domain to the coding sequence
  • AgeI restriction site upstream of stop codon to easily fuse a C-terminal domain to the coding sequence


Source

  • Promoter: fusion promoter between T7 promoter and lac-operator
    • T7 promoter from T7 phage
    • lac-operator from E. coli
  • BBa_K525401 S-layer gene sbpA synthesized, originated in Lysinbacillus sphaericus
  • BBa_J18931 yellow fluorescent protein (YFP) mCitrine
    • from parts.igem
    • was synthesized before it was sent in


References

Kainz B, Steiner K, Möller M, Pum D, Schäffer C, Sleytr UB, Toca-Herrera JL (2010) Absorption, Steady-State Fluorescence, Fluorescence Lifetime, and 2D Self-Assembly Properties of Engineered Fluorescent S-Layer Fusion Proteins of Geobacillus stearothermophilus NRS 2004/3a, [http://pubs.acs.org/doi/abs/10.1021/bm901071b Biomacromolecules 11(1):207-214].

Sleytr UB, Huber C, Ilk N, Pum D, Schuster B, Egelseer EM (2007) S-layers as a tool kit for nanobiotechnological applications, [http://onlinelibrary.wiley.com/doi/10.1111/j.1574-6968.2006.00573.x/full FEMS Microbiol Lett 267(2):131-144].